Conventional bone decalcification is a time-consuming process and is therefore unsuitable for clinical applications and time-limited research projects. Consequently, we compared the effect of four different decalcification solutions applied at three different temperatures, and assessed the rate of decalcification and the implications on tissue morphology and antigenicity of mouse and rat tibiae. Bones were decalcified with 10% ethylenediaminetetraacetic acid (EDTA), 10% formic acid, 5% hydrochloric acid, and 5% nitric acid at 4C, 25C, and 37C. Decalcification in both species was fastest in nitric acid at 37C and slowest in EDTA at 4C. Histological and immunohistochemical staining confirmed that the conventional protocols of EDTA at 4C and 25C remain the best option regarding the quality of tissue preservation. Whereas formic acid at 4C is a good alternative saving about 90% of the decalcification time, hydrochloric and nitric acids should be avoided particularly in case of rat tibia. By contrast, due to their smaller size, mouse tibiae had shorter decalcification times and tolerated higher temperatures and exposure to acids much better. In conclusion, this study demonstrated that depending on the specific research question and sample size, alternative decalcification methods could be used to decrease the time of decalcification while maintaining histological accuracy.

中文翻译：

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小鼠和大鼠胫骨的组织形态和抗原性：比较12种不同的脱钙条件。

传统的骨脱钙是一个耗时的过程，因此不适合临床应用和时间有限的研究项目。因此，我们比较了在三种不同温度下使用的四种不同脱钙溶液的效果，并评估了脱钙的速率及其对小鼠和大鼠胫骨组织形态和抗原性的影响。在4℃，25℃和37℃下用10％乙二胺四乙酸（EDTA），10％甲酸，5％盐酸和5％硝酸将骨头脱钙。在37°C的硝酸中，这两种物质的脱钙速度最快，而在4°C的EDTA中，脱钙速度最慢。组织学和免疫组化染色证实，关于组织保存质量，EDTA的常规方案在4C和25C仍是最佳选择。尽管在4C下使用甲酸是一种很好的替代方法，可节省约90％的脱钙时间，但应避免使用盐酸和硝酸，尤其是在大鼠胫骨的情况下。相比之下，由于其较小的尺寸，小鼠胫骨的脱钙时间较短，并且可以承受较高的温度，并且更好地暴露于酸中。总之，这项研究表明，根据具体的研究问题和样本量，可以使用其他脱钙方法来减少脱钙时间，同时保持组织学准确性。小鼠胫骨的脱钙时间较短，可以承受较高的温度，并且更好地暴露于酸中。总之，这项研究表明，根据具体的研究问题和样本量，可以使用其他脱钙方法来减少脱钙时间，同时保持组织学准确性。小鼠胫骨的脱钙时间较短，可以承受较高的温度，并且更好地暴露于酸中。总之，这项研究表明，根据具体的研究问题和样本量，可以使用其他脱钙方法来减少脱钙时间，同时保持组织学准确性。